Advertisement
1 / 24

1- Making Density Gradients PowerPoint PPT Presentation


  • 418 Views
  • Uploaded on 20-02-2012
  • Presentation posted in: General

1- Making Density Gradients. Pre-formed discontinuous gradients Pre-formed continuous gradients Self-generated gradients. 2- Density Gradient Harvesting. C. A. B. Discontinuous gradient by overlayering. A. B. C. D. Discontinuous gradient by underlayering. density. top. bottom. - PowerPoint PPT Presentation

Download Presentation

1- Making Density Gradients

An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -

Presentation Transcript


1 making density gradients l.jpg

1- Making Density Gradients

  • Pre-formed discontinuous gradients

  • Pre-formed continuous gradients

  • Self-generated gradients

2- Density Gradient Harvesting


Discontinuous gradient by overlayering l.jpg

C

A

B

Discontinuous gradient by overlayering


Discontinuous gradient by underlayering l.jpg

A

B

C

D

Discontinuous gradient by underlayering


Diffusion of a discontinuous gradient l.jpg

density

top

bottom

Diffusion of a discontinuous gradient


Rapid formation of continuous from discontinuous gradient l.jpg

Discontinuous

Continuous

45-60 min

Rapid formation of continuous from discontinuous gradient


Two chamber device for continuous gradients l.jpg

A

B

T

SB

SB

P

M

Two-chamber device for continuous gradients


The gradient master l.jpg

B

The Gradient Master


Gradient master profiles from 10 and 40 iodixanol at 80 and 20 rpm effect of time l.jpg

Density (g/ml)

2 min

1.13

2.5 min

3.0 min

1.11

1.09

1.07

1

3

5

7

9

Fraction Number

Gradient Master profiles from 10% and 40% iodixanol at 80° and 20 rpm: effect of time


Swinging bucket rotor l.jpg

axis of rotation

g

At rest

Spinning

g

Swinging-bucket rotor


Fixed angle and vertical rotors l.jpg

Fixed-angle

Vertical

g

1g

Fixed-angle and vertical rotors


Sedimentation path length of rotors l.jpg

Swinging-bucket

Axis of

rotation

Vertical

Sedimentation path length of rotors


Sample gradient in vertical rotor l.jpg

A

B

Axis of rotation

Sample/gradient in vertical rotor


Self generated gradients l.jpg

time

g

Self-generated gradients

  • At 360,000gav iodixanol molecules will start to sediment


Self generated gradient strategy l.jpg

350,000g

350,000g

1g

1g

Self-generated gradient strategy


Near vertical rotors are best of all l.jpg

Near-vertical rotors are best of all


Iodixanol self generated gradient requirements l.jpg

Iodixanol self-generated gradient requirements

  • Rotors with a short sedimentation path length ~ 17 mm

  • RCF of 180,000-350,000gav


Effect of time on iodixanol self generated gradient in the beckman tln100 at 365 000g l.jpg

Density (g/ml)

15% iodixanol

1.30

1 h

3 h

1.25

20% iodixanol

1.20

1 h

3 h

1.15

1.10

1.05

1.00

1

3

5

7

9

11

13

15

Fraction Number

Effect of time on iodixanol self-generated gradient in the Beckman TLN100 at 365,000g


Effect of rcf and iodixanol in beckman vti65 1 3 h l.jpg

1.3

15% (170,000g)

30% (170,000g)

15% (353,000g)

1.2

30% (353,000g)

Density (g/ml)

1.1

1.0

1

3

5

7

9

11

Fraction Number

Effect of RCF and [iodixanol] in Beckman VTi65.1 (3 h)


Removal of banded material using a syringe l.jpg

Removal of banded material using a syringe


Collecting a band from a sealed tube l.jpg

Collecting a band from a sealed tube


Harvest gradient into equal volume fractions by tube puncture l.jpg

Gilson Fraction

Collector

P

96-well plate

Harvest gradient into equal volume fractions by tube puncture


Upward displacement l.jpg

Perfluorodecalin

Perfluorodecalin

P

P

Upward displacement


Labconco auto densi flow collects from meniscus l.jpg

P

P

P

P

M

B

EP

C

1

2

3

4

Labconco Auto Densi-flow collects from meniscus


Labconco auto densi flow linked to a gilson fc205 fraction collector l.jpg

MB

Labconco Auto Densi-flow linked to a Gilson FC205 Fraction Collector